Volume 10, Issue 3 (6-2007)
J Arak Uni Med Sci 2007, 10(3): 1-7 |
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Abtahi H, Mosayebi G, Soleimanian A H. Expression of recombinant streptolysin O by Escherichia coli. J Arak Uni Med Sci 2007; 10 (3) :1-7
URL: http://jams.arakmu.ac.ir/article-1-49-en.html
URL: http://jams.arakmu.ac.ir/article-1-49-en.html
1- , h_abtahi2@yahoo.co.uk
Abstract: (26428 Views)
Introduction: Streptolysin O (SLO) is an antigenic protein that is secreted by Streptococcus pyogenes. Streptococcal infections are diagnosed with anti streptolysin O. At present, streptolysin O is produced by vectors that have fusion protein. In this study streptolysin O without fusion protein vectors is produced. Materials and Methods: In this experimental study, Streptolysin O gene was amplified by Polymerase chain reaction (PCR) method and subcloned to prokaryotic expression vector pET28a. Escherichia coli BL21-DE3-plySs were transformed with pET28a-SLO and gene expression was induced by IPTG. Then it was purified by Ni-NTA kit. The concentration of SLO was assayed by Bradford method. To confirm recombinant SLO Western Blot was used. Results: The sequencing result was confirmed by Sanger method and was the same as SLO gene. Escherichia coli BL21 (DE3) pLysS was transformed with pET28a-SLO and gene expression was induced by IPTG. The expressed protein was purified by affinity chromatography with Ni-NTA resin. The concentration of purified protein was 100µg/ml. The integrity of product was confirmed by Western Blot analysis using a mouse anti streptolysin O. Conclusion: Data showed that recombinant SLO protein can be produced by pET28a in Escherichia coli. This protein maintains its antigenic effect very well. Therefore, recombinant SLO has same epitopes with natural form of this antigen.
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Received: 2008/09/27
Received: 2008/09/27
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